阿司匹林、二甲双胍预防激素性股骨头坏死的实验研究

发布时间：2018-09-18 18:11

【摘要】：研究背景激素性股骨头坏死是骨科常见的难治性疾病,其疾病进展不可逆,严重影响患者的生活质量,保守治疗效果欠佳。脂质代谢异常、凝血功能障碍、骨髓间充质干细胞(BMSC)的数量和活性下降等因素是引发激素性股骨头坏死的重要原因。目前仍无预防激素性股骨头坏死的特效药物。近年来有研究发现阿司匹林、二甲双胍均具有促进成骨、抑制骨吸收、调节脂质代谢的作用,并能促进骨髓间充质干细胞的成骨分化,因此我们假设阿司匹林和二甲双胍可能具有预防激素性股骨头坏死的作用。研究方法1.应用脂多糖(LPS,200μg/kg,腹腔内注射2次)联合大剂量甲基强的松龙(MPS,40mg/kg,肌肉注射3次)的方法建立大鼠激素性股骨头坏死模型。模型组除股骨头坏死建模外未予特殊处理,实验组在建模的同时每日给予口服l0mg/kg/天的阿司匹林或100mg/kg/天的二甲双胍,持续1.5月/3月后,取大鼠静脉血进行血清骨钙素检测,并处死大鼠,取双侧股骨,行股骨头microCT检查,观察股骨头是否发生坏死,之后将股骨头标本脱钙,行组织病理学检查。2.从1.5月各组大鼠中随机抽取2只进行骨髓间充质干细胞实验。应用全骨髓贴壁法分离培养BMSC,将第1代BMSC进行成骨诱导分化14天后检测其成骨分化相关标志物(碱性磷酸酶、骨钙素、1型胶原、矿化结节)的变化。研究结果1.阿司匹林组1.5月及3月后股骨头坏死率均显著低于模型组(p0.05)。二甲双胍组1.5月后股骨头坏死率低于模型组,但结果无统计学意义(p=0.0995),二甲双胍组3月后股骨头坏死率显著低于模型组(p0.05)。与模型组相比,阿司匹林组和二甲双胍组骨小梁体积、厚度均明显增高。2.采用全骨髓贴壁法可以顺利的分离培养大鼠BMSC。大鼠BMSC在体外可以诱导进行成骨分化。在成骨诱导分化14天后,阿司匹林组BMSC与模型组相比碱性磷酸酶、骨钙素、1型胶原和矿化结节的水平均显著提升(P0.05),二甲双胍组BMSC与模型组相比碱性磷酸酶、]型胶原和矿化结节的水平显著提升(P0.05)。结论1.口服阿司匹林10mg/kg/天持续1.5月或3月均可以显著降低大鼠激素性股骨头坏死的发病率,提高骨小梁体积、厚度等相关参数。2.口服二甲双胍100mg/kg/天持续3月可以显著降低大鼠激素性股骨头坏死的发病率,提高骨小梁体积、厚度等相关参数。3.口服阿司匹林10mg/kg/天或二甲双胍100mg/kg/天持续1.5月,可以增强大鼠激素性股骨头坏死模型中骨髓间充质干细胞的成骨分化能力。
[Abstract]:Background steroid-induced femoral head necrosis is a common refractory disease in orthopedic department. The progress of the disease is irreversible, which seriously affects the quality of life of patients, and the effect of conservative treatment is not good. Abnormal lipid metabolism, dysfunction of coagulation and the decrease of (BMSC) activity of bone marrow mesenchymal stem cells are the important causes of steroid-induced osteonecrosis of femoral head. There is no specific drug available to prevent hormone-induced necrosis of the femoral head. In recent years, aspirin and metformin have been found to promote osteogenesis, inhibit bone resorption, regulate lipid metabolism, and promote osteogenic differentiation of bone marrow mesenchymal stem cells. Therefore, we hypothesized that aspirin and metformin might prevent steroid-induced osteonecrosis of the femoral head. Method 1. The model of steroid-induced femoral head necrosis in rats was established by using lipopolysaccharide (LPS,200 渭 g / kg, intraperitoneal injection twice) and high dose methylprednisolone (MPS,40mg/kg, 3 times intramuscular injection). There was no special treatment in the model group except for the model of femoral head necrosis. The experimental group was given oral aspirin for l0mg/kg/ day and metformin for 100mg/kg/ day daily. After 1.5 months / 3 months, serum osteocalcin was measured in the venous blood of rats. The rats were killed and the femoral head was examined by microCT, then the femoral head was decalcified and histopathological examination was performed. Two bone marrow mesenchymal stem cells were randomly selected from each group for 1.5 months. The changes of osteogenic differentiation related markers (alkaline phosphatase, osteocalcin type 1 collagen, mineralized nodules) were detected 14 days after the first generation BMSC was induced by osteogenic differentiation of BMSC, isolated and cultured by whole bone marrow adherent method, and the changes of osteogenic differentiation markers (alkaline phosphatase, osteocalcin type 1 collagen, mineralized nodules) were detected. Results 1. The necrosis rate of femoral head in aspirin group was significantly lower than that in model group (p0.05). The osteonecrosis rate of femoral head in metformin group was significantly lower than that in model group after 1.5 months (p0.05), but there was no significant difference between metformin group and model group (p0.05). Compared with the model group, the volume and thickness of trabecular bone in aspirin group and metformin group were significantly increased. The method of whole bone marrow adherent can be used to isolate and culture rat BMSC. smoothly. Rat BMSC can induce osteogenic differentiation in vitro. After 14 days of osteogenic differentiation, BMSC in aspirin group was higher than that in model group. The levels of osteocalcin type 1 collagen and mineralized nodules were significantly increased (P0.05), and the levels of BMSC, type 1 collagen and mineralized nodules in metformin group were significantly higher than those in model group (P0.05). Conclusion 1. Oral aspirin 10mg/kg/ for 1.5 months or 3 months could significantly reduce the incidence of steroid-induced femoral head necrosis in rats and increase the trabecular volume and thickness of bone. Oral administration of metformin 100mg/kg/ for 3 months could significantly reduce the incidence of steroid-induced femoral head necrosis and increase the trabecular volume and thickness of bone. Oral administration of aspirin 10mg/kg/ day or metformin 100mg/kg/ day for 1.5 months could enhance the osteogenic differentiation of bone marrow mesenchymal stem cells in steroid-induced femoral head necrosis model in rats.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R681.8

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